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Modulation of the Actin Cytoskeleton via Gelsolin Regulates Vacuolar H+-ATPase Recycling

Modulation of the Actin Cytoskeleton via Gelsolin Regulates Vacuolar H+-ATPase Recycling
Modulation of the Actin Cytoskeleton via Gelsolin Regulates Vacuolar H+-ATPase Recycling
The role of the actin cytoskeleton in regulating membrane protein trafficking is complex and depends on the cell type and protein being examined. Using the epididymis as a model system in which luminal acidification is crucial for sperm maturation and storage, we now report that modulation of the actin cytoskeleton by the calcium-activated actin-capping and -severing protein gelsolin plays a key role in regulating vacuolar H(+)-ATPase (V-ATPase) recycling. Epididymal clear cells contain abundant V-ATPase in their apical pole, and an increase in their cell-surface V-ATPase expression correlates with an increase in luminal proton secretion. We have shown that apical membrane accumulation of V-ATPase is triggered by an elevation in cAMP following activation of bicarbonate-regulated soluble adenylyl cyclase in response to alkaline luminal pH (Pastor-Soler, N., Beaulieu, V., Litvin, T. N., Da Silva, N., Chen, Y., Brown, D., Buck, J., Levin, L. R., and Breton, S. (2003) J. Biol. Chem. 278, 49523-49529). Here, we show that clear cells express high levels of gelsolin, indicating a potential role in the functional activity of these cells. When jasplakinolide was used to overcome the severing action of gelsolin by polymerizing actin, complete inhibition of the alkaline pH- and cAMP-induced apical membrane accumulation of V-ATPase was observed. Conversely, when gelsolin-mediated actin filament elongation was inhibited using a 10-residue peptide (PBP10) derived from the phosphatidylinositol 4,5-bisphosphate-binding region (phosphoinositide-binding domain 2) of gelsolin, significant V-ATPase apical membrane mobilization was induced, even at acidic luminal pH. In contrast, the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl ester) and the phospholipase C inhibitor U-73122 inhibited the alkaline pH-induced V-ATPase apical accumulation. Thus, maintenance of the actin cytoskeleton in a depolymerized state by gelsolin facilitates calcium-dependent apical accumulation of V-ATPase in response to luminal pH alkalinization. Gelsolin is present in other cell types that express the V-ATPase in their plasma membrane and recycling vesicles, including kidney intercalated cells and osteoclasts. Therefore, modulation of the actin cortex by this severing and capping protein may represent a common mechanism by which these cells regulate their rate of proton secretion
0021-9258
8452-8463
Beaulieu, Valerie
563ba5b3-250a-41ad-926e-85708b555500
Da Silva, Nicolas
1e819b5f-b4cf-4606-a919-c8bec5742c24
Pastor-Soler, Nuria
b486bd81-cd99-4984-9fbd-4e732319307c
Brown, Christopher R.
f53b4ced-eb1c-4977-99e7-91203b6e886c
Smith, Peter J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Brown, Dennis
6938f113-e019-4a94-b66a-ae244ae2e6d7
Brenton, Sylvie
71cef2e8-626a-4906-badb-fceb63fa24a1
Beaulieu, Valerie
563ba5b3-250a-41ad-926e-85708b555500
Da Silva, Nicolas
1e819b5f-b4cf-4606-a919-c8bec5742c24
Pastor-Soler, Nuria
b486bd81-cd99-4984-9fbd-4e732319307c
Brown, Christopher R.
f53b4ced-eb1c-4977-99e7-91203b6e886c
Smith, Peter J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Brown, Dennis
6938f113-e019-4a94-b66a-ae244ae2e6d7
Brenton, Sylvie
71cef2e8-626a-4906-badb-fceb63fa24a1

Beaulieu, Valerie, Da Silva, Nicolas, Pastor-Soler, Nuria, Brown, Christopher R., Smith, Peter J.S., Brown, Dennis and Brenton, Sylvie (2004) Modulation of the Actin Cytoskeleton via Gelsolin Regulates Vacuolar H+-ATPase Recycling. The Journal of Biological Chemistry, 280 (9), 8452-8463. (doi:10.1074/jbc.M412750200). (PMID:15591047)

Record type: Article

Abstract

The role of the actin cytoskeleton in regulating membrane protein trafficking is complex and depends on the cell type and protein being examined. Using the epididymis as a model system in which luminal acidification is crucial for sperm maturation and storage, we now report that modulation of the actin cytoskeleton by the calcium-activated actin-capping and -severing protein gelsolin plays a key role in regulating vacuolar H(+)-ATPase (V-ATPase) recycling. Epididymal clear cells contain abundant V-ATPase in their apical pole, and an increase in their cell-surface V-ATPase expression correlates with an increase in luminal proton secretion. We have shown that apical membrane accumulation of V-ATPase is triggered by an elevation in cAMP following activation of bicarbonate-regulated soluble adenylyl cyclase in response to alkaline luminal pH (Pastor-Soler, N., Beaulieu, V., Litvin, T. N., Da Silva, N., Chen, Y., Brown, D., Buck, J., Levin, L. R., and Breton, S. (2003) J. Biol. Chem. 278, 49523-49529). Here, we show that clear cells express high levels of gelsolin, indicating a potential role in the functional activity of these cells. When jasplakinolide was used to overcome the severing action of gelsolin by polymerizing actin, complete inhibition of the alkaline pH- and cAMP-induced apical membrane accumulation of V-ATPase was observed. Conversely, when gelsolin-mediated actin filament elongation was inhibited using a 10-residue peptide (PBP10) derived from the phosphatidylinositol 4,5-bisphosphate-binding region (phosphoinositide-binding domain 2) of gelsolin, significant V-ATPase apical membrane mobilization was induced, even at acidic luminal pH. In contrast, the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl ester) and the phospholipase C inhibitor U-73122 inhibited the alkaline pH-induced V-ATPase apical accumulation. Thus, maintenance of the actin cytoskeleton in a depolymerized state by gelsolin facilitates calcium-dependent apical accumulation of V-ATPase in response to luminal pH alkalinization. Gelsolin is present in other cell types that express the V-ATPase in their plasma membrane and recycling vesicles, including kidney intercalated cells and osteoclasts. Therefore, modulation of the actin cortex by this severing and capping protein may represent a common mechanism by which these cells regulate their rate of proton secretion

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Published date: March 2004
Organisations: University of Southampton

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Local EPrints ID: 188763
URI: https://eprints.soton.ac.uk/id/eprint/188763
ISSN: 0021-9258
PURE UUID: bcd0cd91-370b-4116-8926-347750465097
ORCID for Peter J.S. Smith: ORCID iD orcid.org/0000-0003-4400-6853

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Date deposited: 06 Jun 2011 09:15
Last modified: 06 Jun 2018 12:31

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Contributors

Author: Valerie Beaulieu
Author: Nicolas Da Silva
Author: Nuria Pastor-Soler
Author: Christopher R. Brown
Author: Dennis Brown
Author: Sylvie Brenton

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